• Title/Summary/Keyword: conidial production

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Menadione Sodium Bisulfite-Protected Tomato Leaves against Grey Mould via Antifungal Activity and Enhanced Plant Immunity

  • Jo, Youn Sook;Park, Hye Bin;Kim, Ji Yun;Choi, Seong Min;Lee, Da Sol;Kim, Do Hoon;Lee, Young Hee;Park, Chang-Jin;Jeun, Yong-Chull;Hong, Jeum Kyu
    • The Plant Pathology Journal
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    • v.36 no.4
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    • pp.335-345
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    • 2020
  • Tomato grey mould has been one of the destructive fungal diseases during tomato production. Ten mM of menadione sodium bisulfite (MSB) was applied to tomato plants for eco-friendly control of the grey mould. MSB-reduced tomato grey mould in the 3rd true leaves was prolonged at least 7 days prior to the fungal inoculation of two inoculum densities (2 × 104 and 2 × 105 conidia/ml) of Botrytis cinerea. Protection efficacy was significantly higher in the leaves inoculated with the lower disease pressure of conidial suspension compared to the higher one. MSB-pretreatment was not effective to arrest oxalic acid-triggered necrosis on tomato leaves. Plant cell death and hydrogen peroxide accumulation were restricted in necrotic lesions of the B. cinereainoculated leaves by the MSB-pretreatment. Decreased conidia number and germ-tube elongation of B. cinerea were found at 10 h, and mycelial growth was also impeded at 24 h on the MSB-pretreated leaves. MSB-mediated disease suppressions were found in cotyledons and different positions (1st to 5th) of true leaves inoculated with the lower conidial suspension, but only 1st to 3rd true leaves showed decreases in lesion sizes by the higher inoculum density. Increasing MSB-pretreatment times more efficiently decreased the lesion size by the higher disease pressure. MSB led to inducible expressions of defence-related genes SlPR1a, SlPR1b, SlPIN2, SlACO1, SlChi3, and SlChi9 in tomato leaves prior to B. cinerea infection. These results suggest that MSB pretreatment can be a promising alternative to chemical fungicides for environment-friendly management of tomato grey mould.

Culture Method of Spore for Entomopathogenic Fungus Using Natural Zeolite Ceramic Ball (천연제오라이트 세라믹볼을 이용한 곤충병원성 곰팡이 포자 생산 방법)

  • Lee, Jung-Bok;Kim, Beaum-Soo;Joo, Woo-Hong;Kwon, Gi-Seok
    • Korean Journal of Environmental Agriculture
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    • v.35 no.1
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    • pp.72-78
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    • 2016
  • BACKGROUND: Entomopathogenic fungi have been studied to develop for biological control agents as an alternative to chemical control agents in insect pest management. This investigated to determine the optimal culture conditions in ceramic balls for maximal sporulation of entomopathogenic fungi Beauveria bassiana M130 by use rice bran extract.METHODS AND RESULTS: METHODS AND RESULTS: A culture of entomopathogenic fungi for 12day on rice bran extract(1:8, w/v) incubated in ceramic matrix at 28℃. Natural zeolite ceramic ball was high production of 4.2×108 conidial/mL. The culture condition optimized initial pH, temperature, rice bran extract concentration, adhesives substance and concentration of NaCl, respectively. The high production of spore optimal conditions were temperature 28℃, initial pH 3, rice bran extract 3 mL, starch 33 g, 5 % NaCl and sopre suspension 7 mL, respectively.CONCLUSION: This study was carried out for the mass production of entomopathogenic fungi conidia recover rate 65% in matrix of natural zeolite ceramic ball, and to develop ingredient-used formulation of Beauveria bassiana M130 conidia for biological control agents.

Ultrastructure of the Cell Wall of a Null Pigmentation Mutant, npgA1, in Aspergillus nidulans

  • Chung, Yun-Shin;Kim, Jung-Mi;Han, Dong-Min;Chae, Keon-Sang;Jahng, Kwang-Yeop
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.224-231
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    • 2003
  • The null pigmentation mutant (npgA1) of Aspergillus nidulans was previously characterized by its production of no pigment at any stage of its life cycle, its reduction in hyphal branching, and its delay in the asexual spore development. The chemical composition of the cell wall was also altered in npgA1 mutants that became more sensitive to Novozyme 234$\^$TM/, which is possibly due to a structural defect in the cell wall. To investigate the effects of the cell wall structure on these pleiomorphic phenomena, we examined the ultrastructure of the cell wall in the npgA1 mutant (WX17). Scanning electron micrographs (SEM) showed that after being cultured for six days, the outermost layer of the conidial wall of WX17 peeled off. Although this phenotype suggested that the cell wall structure in WX17 may be modified, examination using TEM of the fine structure of cross-sectioned hyphal wall of WX17 did not show any differences from that of FGSC4. However, staining for carbohydrates of wall layers showed that the electron-translucent layer of the cell wall was missing in WX17. In addition, the outermost layer H1 of the hyphal wall was also absent in WX17. The ultrastructural observation and cytochemical analysis of cell walls suggested that the pigmentation defect in WX17 may be attributed to the lack of a layer in the cell wall.

Development of Lipase Hyper-producing Strain from Hybrids between Aspergillus niger and Penicillium chrysogenum by Nuclear Transfer (핵전이에 의한 Aspergillus niger와 Penicillium chrysogenum의 잡종에서의 Lipase 고생산균주의 개발)

  • 양영기;문명님;이윤희;임채영
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.31-37
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    • 1997
  • Intergeneric hybrids between Aspergillus niger and Perricillium ch~y.sop~um(Tyr ), hyperlipolytic enzyne-producing fungi, were obtained by nuclear transfer technique:. Optimal conditions for formation of intergeneric hybrids were investigated. Maximum production of protoplasts were obtainrd by 1% Novozym 234 at $30^{\circ}C$ for 3 hrs and the most effective osmotic stabilizers for the isolation of protoplasts were 0.6 M KC]. Frequencies of hybrid formation by nuclear transfer were $1.3{\times}$10^{-3}$$ $-3.8{\times}$10^{-3}$$. From the chervation of genetic stability, conidial size, DNA content, ;md nuclear stain, it was suggested that their karyotypes are aneuploid. The hybrids showed 1.4-2.2 fold higher lipase activities than parental strains. It was strongly supported by results of this study that nuclear transfer technique is much more efficient in the formation of intergeneric hybrids than protoplast fusion and is very useful for the improvement of strains.

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Intergeneric Hybrid Constructed by Nuclear Transfer of Saccharomycopsis into Saccharomyces (핵전이를 이용한 Saccharomycopsis 속과 Saccharomyces 속간의 잡종형성)

  • Yang, Young-Ki;Lim, Chae-Young;Kang, Hee-Kyoung;Moon, Myeng-Nim;Rhee, Young-Ha
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.399-405
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    • 1999
  • Intergeneric hybrids between Saccharomyccopsis fiburigera KCTC 7393 and Saccharomyces cerevisiae KCTC 7049 (tyr-, ura-) were obtained by nuclear transfer technique. Nuclei isolated from the wild type S. fiburigera strain were transfered into auxotrophic S. cerevisiae mutants and new strains showing an increased starch degrading capability were selected. Maximum production of protoplasts was obtained from the treatment with 0.1 % Novozym 234 at $30^{\circ}C$ for 90 min, and most effective osmotic stabilizer for the isolation of protoplasts was 0.6 M KCl at pH 5.8. The frequency of protoplast regeneration was 14.64% under the conditions. Genectic stability, conidial size, DNA content, and nuclear stain suggested that the fusants were aneuploidy. The specific activity of ${\alpha}-amylase$ was observed to increase about $1.2{\sim}1.9$ folds.

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A Short-chain Dehydrogenase/reductase Gene is Required for Infection-related Development and Pathogenicity in Magnaporthe oryzae

  • Kwon, Min-Jung;Kim, Kyoung-Su;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.8-16
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    • 2010
  • The phytopathogenic fungus Magnaporthe oryzae is a major limiting factor in rice production. To understand the genetic basis of M. oryzae pathogenic development, we previously analyzed a library of T-DNA insertional mutants of M. oryzae, and identified ATMT0879A1 as one of the pathogenicity-defective mutants. Molecular analyses and database searches revealed that a single TDNA insertion in ATMT0879A1 resulted in functional interference with an annotated gene, MGG00056, which encodes a short-chain dehydrogenase/reductase (SDR). The mutant and annotated gene were designated as $MoSDR1^{T-DNA}$ and MoSDR1, respectively. Like other SDR family members, MoSDR1 possesses both a cofactor-binding motif and a catalytic site. The expression pattern of MoSDR1 suggests that the gene is associated with pathogenicity and plays an important role in M. oryzae development. To understand the roles of MoSDR1, the deletion mutant ${\Delta}Mosdr1$ for the gene was obtained via homology-dependent gene replacement. As expected, ${\Delta}Mosdr1$ was nonpathogenic; moreover, the mutant displayed pleiotropic defects in conidiation, conidial germination, appressorium formation, penetration, and growth inside host tissues. These results suggest that MoSDR1 functions as a key metabolic enzyme in the regulation of development and pathogenicity in M. oryzae.

Identification of Genes Encoding Heat Shock Protein 40 Family and the Functional Characterization of Two Hsp40s, MHF16 and MHF21, in Magnaporthe oryzae

  • Yi, Mi-Hwa;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.131-142
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    • 2008
  • Magnaporthe oryzae, the causal agent of the rice blast disease, poses a worldwide threat to stable rice production. The large-scale functional characterization of genes controlling the pathogenicity of M. oryzae is currently under way, but little is known about heat shock protein 40 (Hsp40) function in the rice blast fungus or any other filamentous plant pathogen. We identified 25 genes encoding putative Hsp40s in the genome of M. oryzae using a bioinformatic approach, which we designated M. oryzae heat shock protein forty (MHF 1-25). To elucidate the roles of these genes, we characterized the functions of MHF16 and MHF21, which encode type ill and type n Hsp40 proteins, respectively. MHF16 and MHF21 expression was not significantly induced by heat shock, but it was down-regulated by cold shock. Knockout mutants of these genes $({\Delta}$mhf16 and ${\Delta}$mhf21) were viable, but conidiation was severely reduced. Moreover, sectoring was observed in the ${\Delta}mhf16$ mutant when it was grown on oatmeal agar medium. Conidial germination, appressorium formation, and pathogenicity in rice were not significantly affected in the mutants. The defects in conidiation and colony morphology were fully complemented by reintroduction of wild type MHF16 and MHF21 alleles, respectively. These data indicate that MHF16 and MHF21 play important roles in conidiation in the rice blast fungus.

Antagonistic Activities of Bacillus spp. Strains Isolated from Tidal Flat Sediment Towards Anthracnose Pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea

  • Han, Joon-Hee;Shim, Hongsik;Shin, Jong-Hwan;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.165-175
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    • 2015
  • Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

Biocontrol Characteristics of Bacillus Species in Suppressing Stem Rot of Grafted Cactus Caused by Bipolaris cactivora

  • Bae, Sooil;Kim, Sang Gyu;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.42-51
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    • 2013
  • One of the most important limiting factors for the production of the grafted cactus in Korea is the qualitative and quantitative yield loss derived from stem rots especially caused by Bipolaris cactivora. This study is aimed to develop microbial control agents useful for the control of the bipolaris stem rot. Two bacteria (GA1-23 and GA4-4) selected out of 943 microbial isolates because of their strong antibiotic activity against B. cactivora were identified as Bacillus subtilis and B. amyloliquefaciens, respectively, by the cultural characteristics, Biolog program and 16S rRNA sequencing analyses. Both bacterial isolates significantly inhibited the conidial germination and mycelial growth of the pathogen with no significant difference between the two, of which the inhibitory efficacies varied depending on the cultural conditions such as temperature, nutritional compositions and concentrations. Light and electron microscopy of the pathogen treated with the bacterial isolates showed the inhibition of spore germination with initial malformation of germ tubes and later formation of circle-like vesicles with no hyphal growth and hyphal disruption sometimes accompanied by hyphal swellings and shrinkages adjacent to the bacteria, suggesting their antibiotic mode of antagonistic activity. Control efficacy of B. subtilis GA1-23 and B. amyloliquefaciens GA4-4 on the cactus stem rot were not as high as but comparable to that of fungicide difenoconazole when they were treated simultaneously at the time of pathogen inoculation. All of these results suggest the two bacterial isolates have a good potential to be developed as biocontrol agents for the bipolaris stem rot of the grafted cactus.

The Small GTPase CsRAC1 Is Important for Fungal Development and Pepper Anthracnose in Colletotrichum scovillei

  • Lee, Noh-Hyun;Fu, Teng;Shin, Jong-Hwan;Song, Yong-Won;Jang, Dong-Cheol;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.607-618
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    • 2021
  • The pepper anthracnose fungus, Colletotrichum scovillei, causes severe losses of pepper fruit production in the tropical and temperate zones. RAC1 is a highly conserved small GTP-binding protein in the Rho GT-Pase family. This protein has been demonstrated to play a role in fungal development, and pathogenicity in several plant pathogenic fungi. However, the functional roles of RAC1 are not characterized in C. scovillei causing anthracnose on pepper fruits. Here, we generated a deletion mutant (𝜟Csrac1) via homologous recombination to investigate the functional roles of CsRAC1. The 𝜟Csrac1 showed pleiotropic defects in fungal growth and developments, including vegetative growth, conidiogenesis, conidial germination and appressorium formation, compared to wild-type. Although 𝜟Csrac1 was able to develop appressoria, it failed to differentiate appressorium pegs. However, 𝜟Csrac1 still caused anthracnose disease with significantly reduced rate on wounded pepper fruits. Further analyses revealed that 𝜟Csrac1 was defective in tolerance to oxidative stress and suppression of host-defense genes. Taken together, our results suggest that CsRAC1 plays essential roles in fungal development and pathogenicity in C. scovilleipepper fruit pathosystem.